Dissertation > Excellent graduate degree dissertation topics show

Effect of Liposomal Transfection of UCH-L1 siRNA on Proliferation and Apoptosis of H157 Cell

Author: QuXiaoJie
Tutor: WangYanFu
School: Dalian Medical University
Course: Respiration within the science
Keywords: Ubiquitin carboxy-terminal hydrolase -1 SiRNA Lung cancer
CLC: R734.2
Type: Master's thesis
Year: 2010
Downloads: 105
Quote: 0
Read: Download Dissertation

Abstract

Objective: Lung cancer is the world's highest incidence and mortality of malignant tumors, 60% of the clinical diagnosis of lung cancer in advanced stage, despite surgery, radiotherapy and chemotherapy treatment, reasons for the transfer and multi-drug resistant non-small cell lung cancer (non-small cell lung cancer, NSCLC) 5-year survival rate is still lower than 10%, less than 3% of small cell lung cancer (small cell lung cancer, SCLC). There is an urgent need to find new diagnostic indicators and effective treatment methods. Tumorigenesis is a multi-factor, multi-stage, multiple genes involved in complex and long-term development process, in which the gene change is the most critical factor. Therefore, how to clarify from the gene's point of lung cancer, the development mechanism and to find effective therapeutic target to become the present study the difficult and hot research has opened up new avenues of development and wide application of modern molecular biology techniques in the field. Ubiquitin carboxyl-terminal hydrolase -1 (Ubiquitin C terminal hydrolase-L1, UCH-L1) is a recent study more of carcinoid gene in a variety of malignancies, such as esophageal cancer, lung cancer, breast cancer in both abnormal expression and increased activity. UCH-L1 molecular weight of 27 KD, belonging to the the ubiquitin peptide carboxyl-terminal hydrolases family, in the cell proteolytic pathways regulating cellular processes including cell division cycle and death. It is widely expressed in the various stages of neuronal differentiation, the markers of a particular organization of the neuroendocrine system. UCH-L 1-mediated targeted cell protein ubiquitin - proteasome system, may be an important mechanism of regulation of cell cycle genes by causing cell cycle protein deubiquitination increased to promote the growth and reproduction of undifferentiated somatic disorder may induce expression of UCH-L1 gene can affect the cell division cycle. Therefore UCH-L1 in regulating the expression of cell cycle proteins has an important role in malignancy development and prognosis. The purpose of this study is to explore through liposome the UCH-L1siRNA transfected human lung adenocarcinoma H157 cells, lung adenocarcinoma cell research after transfection morphological changes H157, H157 cells detected by flow cytometry apoptosis and cell cycle changes H157 cells detected RNA levels and protein levels after transfection, apoptosis and proliferation. : Synthetic inhibit UCH-L 1 gene siRNA fragments by liposome transfected into human lung adenocarcinoma H157 cells, the morphological changes observed under the microscope; flow cytometry cell cycle and apoptosis Index; RT-PCR was used to detect UCH-L1 mRNA expression; Western blot analysis detected UCH-L1 protein expression. Results: inverted microscope morphological changes of transfected cells: before transfection, cells adherent better, was flat polygonal middle of round nucleus, the nucleus from one to multiple nucleoli, 48 hours after transfection, cells volume shrinkage, poor adherence, cell processes disappear. Flow cytometry cell cycle and apoptosis: The results showed that UCH-L1 siRNA transfected cells was significantly arrest at the G2 / M phase and apoptosis rate of the three groups were UCH-L1 siRNA transfection group (28.06 ± 1.58%), and blank control group (0.45 ± 0.09)%, negative control group (0.52 ± 0.07)%, UCH-L1 siRNA can significantly inhibit UCH-L1, induction of apoptosis. RT-PCR, UCH-L1 mRNA expression, and results show that the UCH-Ll siRNA effect in H157 cells after 48h UCH-L1 mRNA expression was significantly lowered, and blank control group, negative control group of UCH-L 1mRNA in did not change significantly. That UCH-L1 siRNA can induce specific degradation of UCH-L1 mRNA. Western blot analysis of UCH-L1 protein expression, the results show that the role of UCH-L1 siRNA in H157 cells after 48 h the UCH-L1 protein expression was significantly down, and blank control group, no significant change in the negative control group of UCH-L 1 protein levels. Show that the UCH-L1siRNA can induce UCH-L1 protein specific degradation. Conclusion: The application of RNA interference targeting inhibition of UCH-L1 gene can significantly inhibit the proliferation of H157 cells and promote apoptosis. UCH-L 1 may become a new target for gene therapy of lung cancer.

Effect of Liposomal Transfection of UCH-L1 siRNA on Proliferation and Apoptosis of H157 Cell

Abstract

Related Dissertations